Department of Applied Clinical Sciences and Biotechnology, University of L'Aquila, 67100 L'Aquila, Italy.
Department of Oncology and Hematology, Ospedali Riuniti Marche Nord, 61121 Pesaro, Italy.
Int J Mol Sci. 2020 Jul 7;21(13):4813. doi: 10.3390/ijms21134813.
Circulating tumour cells (CTCs) from liquid biopsies are under current investigation in several cancers, including epithelial ovarian cancer (EOC) but face significant drawbacks in terms of non-standardised methodology, low viable cell numbers and accuracy of CTC identification. In this pilot study, we report that chemosensitivity assays using liquid biopsy-derived metastatic EOC CTCs, from 10 patients, nine with stage IIIC and one with stage IV disease, in progression after systemic chemotherapy, submitted for hypoxic isolated abdominal perfusion (HAP), are both feasible and useful in predicting response to therapy. Viable metastatic EOC CTCs (>5 cells/mL for all 10 blood samples), enriched by transient culture and identified by reverse transcription polymerase chain reaction (RT-PCR) and indirect immunofluorescence (IF), were subjected to flow cytometry-based Annexin V-PE assays for chemosensitivity to several chemotherapeutic agents and by RT-PCR for tumour gene expression profiling. Using a cut-off value of >80% cell death, CTC chemosensitivity tests were predictive of patient RECIST 1.1 responses to HAP therapy associated with 100% sensitivity, 50% specificity, 33% positive predictive, 100% negative predictive and 60% accuracy values. We propose that the methodology employed in this study is feasible and has the potential to predict response to therapy, setting the stage for a larger study.
循环肿瘤细胞（CTCs）的液体活检目前正在多种癌症中进行研究，包括上皮性卵巢癌（EOC），但在方法标准化、活细胞数量低和 CTC 鉴定准确性方面存在显著缺陷。在这项初步研究中，我们报告了使用液体活检衍生的转移性 EOC CTC 进行化疗敏感性测定的情况，这些 CTC 来自 10 名患者，其中 9 名处于 III C 期，1 名处于 IV 期疾病，在全身化疗后进展，进行缺氧孤立腹腔灌注（HAP）治疗，这在预测治疗反应方面既可行又有用。通过瞬时培养富集的有活力的转移性 EOC CTC（所有 10 个血液样本均>5 个细胞/mL），通过逆转录聚合酶链反应（RT-PCR）和间接免疫荧光（IF）鉴定，进行基于流式细胞术的 Annexin V-PE 化疗敏感性测定，并通过 RT-PCR 进行肿瘤基因表达谱分析。使用>80%细胞死亡的截止值，CTC 化疗敏感性测试可预测患者对 HAP 治疗的 RECIST 1.1 反应，其敏感性为 100%，特异性为 50%，阳性预测值为 33%，阴性预测值为 100%，准确性为 60%。我们提出，本研究中采用的方法是可行的，并且有可能预测治疗反应，为更大规模的研究奠定了基础。
